Fig. 5.
Scatterplots and western blot analysis of study 2. Scatterplot displaying the peptide-to-total full-length Ng ratio ×1000 for Ng53–78. #Sample ratio is > 150,000 (a). Scatterplot displaying total full-length Ng concentration measured by HI–MS (b). WB using the monoclonal antibody Ng36 (left blot) or no primary antibody (right blot). Briefly, samples were relatively quantified in WB by including a standard curve of recombinant Ng-MYC-DDK protein on each gel. Lanes 1–5: standard curve of Ng–Myc–DKK fusion protein ranging from 25 to 1.56 ng. Lane 6: quality control (QC) brain tissue sample. Lanes 7–9: patient samples from study 2. Lane 10: QC tissue sample. Lane 11: Ng–Myc–DKK fusion protein 12.5 ng. Lanes 10 and 11 were used as negative controls (c). Scatterplot displaying results from WB analysis. The y-axis displays the Ng concentration in ng/mg total protein in each sample (d). Scatterplot displaying ELISA results. The y-axis displays the Ng concentration in ng/mg total protein in each sample (e). The data presented are median and interquartile ranges. Comparisons between groups were performed using Kruskal–Wallis test with data adjusted for post-mortem delay, followed by pairwise Mann–Whitney U tests if significant